Traditional Chinese medicine bulbus fritillariae cirrhosae DNA extraction reagent and extraction method

An extraction method and fritillary technology, which are applied in the field of traditional Chinese medicine fritillary DNA extraction reagents and extraction, can solve problems such as poor DNA integrity, and achieve the effects of optimizing treatment reagents, reducing pollution, and achieving good removal effects.

Pending Publication Date: 2020-08-25
康美华大基因技术有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the kit method is simple and time-consuming,

Method used

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  • Traditional Chinese medicine bulbus fritillariae cirrhosae DNA extraction reagent and extraction method
  • Traditional Chinese medicine bulbus fritillariae cirrhosae DNA extraction reagent and extraction method
  • Traditional Chinese medicine bulbus fritillariae cirrhosae DNA extraction reagent and extraction method

Examples

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Effect test

Embodiment 1

[0036]A traditional Chinese medicine Fritillaria DNA extraction reagent, including nuclear lysate, CTAB buffer, TE Buffer, wherein:

[0037] Nuclear lysate includes: 1mol / L pH 8.0 Tris-HCl 100mL, 0.5mol / L pH 8.0EDTA 10mL, NaCl 14.6g, PVP-40 20g, β-mercaptoethanol 2ml, ddH 2 O was fixed to 1000mL;

[0038] CTAB buffer solution includes: CTAB 30g, NaCl 81.9g, 0.5mol / L pH 8.0EDTA 40mL, 1mol / L pH8.0 Tris-HCl 100mL, PVP-40 20g, β-mercaptoethanol 2ml, ddH 2 O was fixed to 1000mL;

[0039] TE Buffer includes: 1M pH8.0 Tris-HCl Buffer 10mL, 0.5M pH8.0 EDTA 2mL, ddH 2 O was adjusted to 1000mL.

[0040] The Chinese medicine Fritillaria DNA of the present embodiment is extracted by the following method:

[0041] (1) Take 100 mg of the ground sample, add 800 μL of nuclear lysate to each sample tube, vigorously shake and mix for 5 minutes, centrifuge at 12000 r / min for 3 minutes, and discard the supernatant;

[0042] (2) Repeat step (1) 1 to 2 times until the supernatant is not viscou...

Embodiment 2

[0053] A traditional Chinese medicine Fritillaria DNA extraction reagent, including nuclear lysate, CTAB buffer, TE Buffer, wherein:

[0054] Nuclear lysate includes: 1mol / L pH 8.0 Tris-HCl 90mL, 0.5mol / L pH 8.0EDTA 8mL, NaCl 13g, PVP-40 18g, β-mercaptoethanol 1.5ml, ddH 2 O was fixed to 1000mL;

[0055] CTAB buffer solution includes: CTAB 28g, NaCl 75g, 0.5mol / L pH 8.0EDTA 35mL, 1mol / L pH8.0 Tris-HCl 95mL, PVP-40 18g, β-mercaptoethanol 1.5ml, ddH 2 O was fixed to 1000mL;

[0056] TE Buffer includes: 1M pH8.0 Tris-HCl Buffer 10mL, 0.5M pH8.0 EDTA 2mL, ddH 2 O was adjusted to 1000mL.

[0057] The DNA extraction method of the traditional Chinese medicine Fritillaria in this example is the same as that in Example 1, and will not be described in detail here.

Embodiment 3

[0059] A traditional Chinese medicine Fritillaria DNA extraction reagent, including nuclear lysate, CTAB buffer, TE Buffer, wherein:

[0060] Nuclear lysate includes: 1mol / L pH 8.0 Tris-HCl 110mL, 0.5mol / L pH 8.0EDTA 12mL, NaCl 16g, PVP-40 22g, β-mercaptoethanol 2.5ml, ddH 2 O was fixed to 1000mL;

[0061] CTAB buffer solution includes: CTAB 32g, NaCl 85g, 0.5mol / L pH 8.0EDTA 45mL, 1mol / L pH8.0 Tris-HCl 105mL, PVP-40 22g, β-mercaptoethanol 2.5ml, ddH 2 O was fixed to 1000mL;

[0062] TE Buffer includes: 1M pH8.0 Tris-HCl Buffer 10mL, 0.5M pH8.0 EDTA 2mL, ddH2O to 1000mL.

[0063] The DNA extraction method of the traditional Chinese medicine Fritillaria in this example is the same as that in Example 1, and will not be described in detail here.

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Abstract

The invention discloses a traditional Chinese medicine bulbus fritillariae cirrhosae DNA extraction reagent. The extraction reagent comprises a nuclear lysis solution, a CTAB buffer solution and a TEBuffer, wherein the nuclear lysis solution comprises Tris-HCl, EDTA, NaCl, PVP-40 and beta-mercaptoethanol, and is similar to the CTAB buffer solution in component. The DNA extraction reagent is suitable for extraction of traditional Chinese medicine bulbus fritillariae cirrhosae DNA, the removal rate of secondary metabolites is high, and the bulbus fritillariae cirrhosae DNA integrity is high. The invention further provides a traditional Chinese medicine bulbus fritillariae cirrhosae DNA extraction method. By adopting the extraction reagent, based on an existing CTAB method, improvement of anextraction reagent formula and optimization of extraction steps are carried out, so that high-purity and high-quality bulbus fritillariae cirrhosae total DNA can be obtained; and the extraction method is simple, easy to control and low in pollution.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a DNA extraction reagent and an extraction method of a traditional Chinese medicine Fritillaria japonicus. Background technique [0002] At present, molecular biology technology has been widely used in the identification of Chinese herbal medicines, among which the extraction and purification of DNA is an important basis for the study of Chinese herbal medicines. Quickly and effectively extracting a large amount of high-quality DNA is the key to the implementation of molecular markers, gene An important prerequisite for library construction, gene isolation, genetic transformation and molecular identification. However, after most Chinese herbal medicines are processed, the DNA contained in them may be degraded to varying degrees, which is not conducive to maintaining the integrity of DNA; secondly, the secondary metabolites contained in Chinese herbal medicines are diffi...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 胡悦许冬瑾蒙洁妙
Owner 康美华大基因技术有限公司
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